Immortalized Human Primary Coronary Artery Endothelial Cells

$3,000.00

Immortalized Human Primary Coronary Artery Endothelial Cells (HCAECs) are vascular cells derived from coronary arteries and genetically modified to maintain proliferative capacity while retaining key endothelial characteristics. These cells exhibit typical cobblestone morphology and express standard endothelial markers including CD31, von Willebrand Factor (vWF), and VE-cadherin, while testing negative for smooth muscle (α-SMA) and fibroblast (TE-7) contaminants. Certified free of mycoplasma, bacteria, fungi, and endotoxins, the immortalized HCAECs demonstrate stable growth for 30+ passages in specialized endothelial cell medium when cultured on gelatin-coated surfaces at 37°C/5% CO₂. Particularly valuable for cardiovascular research, they are ideal for studying atherosclerosis, angiogenesis, vascular inflammation, and endothelial barrier function. Supplied cryopreserved at passage 4 with ≥1×10⁶ viable cells/vial, these coronary artery endothelial cells maintain functional properties including LDL uptake, nitric oxide production, and leukocyte adhesion molecule expression (VCAM-1, ICAM-1) in response to inflammatory stimuli. Their extended lifespan and preserved vascular functionality make them excellent for both acute mechanistic studies and chronic disease modeling of coronary artery

Immortalized Human Primary Coronary Artery Endothelial Cells (HCAECs) are vascular cells derived from coronary arteries and genetically modified to maintain proliferative capacity while retaining key endothelial characteristics. These cells exhibit typical cobblestone morphology and express standard endothelial markers including CD31, von Willebrand Factor (vWF), and VE-cadherin, while testing negative for smooth muscle (α-SMA) and fibroblast (TE-7) contaminants. Certified free of mycoplasma, bacteria, fungi, and endotoxins, the immortalized HCAECs demonstrate stable growth for 30+ passages in specialized endothelial cell medium when cultured on gelatin-coated surfaces at 37°C/5% CO₂. Particularly valuable for cardiovascular research, they are ideal for studying atherosclerosis, angiogenesis, vascular inflammation, and endothelial barrier function. Supplied cryopreserved at passage 4 with ≥1×10⁶ viable cells/vial, these coronary artery endothelial cells maintain functional properties including LDL uptake, nitric oxide production, and leukocyte adhesion molecule expression (VCAM-1, ICAM-1) in response to inflammatory stimuli. Their extended lifespan and preserved vascular functionality make them excellent for both acute mechanistic studies and chronic disease modeling of coronary artery

Materials Required

  • Immortalized Human Primary Coronary Artery Endothelial Cells

  • Complete Endothelial Cell Growth Medium

  • Tissue culture flasks or plates (T25 or multiwell) pre-coated with Alphabiocoat

  • Phosphate-buffered saline (PBS), sterile

  • Cell detachment solution

  • Neutralizing solution

  • 37°C humidified incubator with 5% CO₂

  • Centrifuge (for cell passaging)

  • Hemocytometer or automated cell counter

  • Cryopreservation medium

  • Liquid nitrogen storage system

Cell Culture Protocol

1. Thawing Frozen Cells

  • Coat Flask: Prepare a T25 flask by adding 1 mL Alphabiocoat. Incubate for 30 minutes at 37°C or overnight at 4°C. Aspirate excess before use.

  • Rinse: Wash the coated flask with sterile 1× PBS.

  • Warm Medium: Bring complete endothelial growth medium to 37°C.

  • Thaw Cells: Quickly thaw the cryovial in a 37°C water bath (1–2 minutes).

  • Dilute Slowly: Transfer contents to a 15 mL tube. Add 5 mL of pre-warmed medium dropwise to minimize osmotic shock.

  • Centrifuge: Spin at 200 × g for 5 minutes.

  • Resuspend & Plate: Remove the supernatant, resuspend the cell pellet in fresh medium, and seed into the prepared flask.

  • Incubate: Place in a 37°C, 5% CO₂ incubator.

2. Routine Maintenance & Passaging

  • Daily Monitoring: Examine cells under a microscope. Healthy cells display a cobblestone-like morphology.

  • Medium Changes: Replace the medium every 2–3 days to prevent waste buildup.

  • Passaging at 80–90% Confluence:

    • Aspirate the medium and gently rinse with PBS.

    • Add cell detachment solution (1 mL for T25, 2–3 mL for larger flasks) and incubate for 2–3 minutes at 37°C.

    • Check under a microscope to confirm detachment.

    • Add 2–3 mL neutralizing solution to stop enzymatic activity.

    • Gently pipette to obtain a single-cell suspension.

    • Optional: Centrifuge at 200 × g for 5 minutes if further washing is needed.

    • Resuspend and replate at a 1:2 to 1:4 split ratio into fresh Alphabiocoat-coated flasks.

3. Cryopreservation

  • Harvest Cells: Follow the passaging procedure to collect cells.

  • Cell Count: Adjust to a final concentration of 1–5 × 10⁶ cells/mL in cryopreservation medium.

  • Aliquot: Dispense 1 mL per cryovial.

  • Freeze Gradually: Place vials in a controlled-rate freezing container or isopropanol freezing container at –80°C overnight.

  • Storage: Transfer vials to liquid nitrogen for long-term storage.

4. Key Considerations & Troubleshooting

  • ✅ Morphology Check: Cells should form a cobblestone monolayer. Elongated or spindle-like cells may indicate contamination or stress.

  • ✅ Avoid Over-confluence: Prolonged confluence may cause detachment or death.

  • ✅ Low Passage Use: For optimal results, use cells within 20–30 passages.

  • ✅ Prevent Contamination: Use sterile technique and antibiotics; endothelial cells are particularly sensitive to microbial contamination.

Expected Results

  • Initial Adhesion: Within 4–6 hours post-plating

  • Proliferation: Noticeable within 24–48 hours

  • Doubling Time: Approximately 24–36 hours, depending on cell line and growth conditions